Biochemical Journal

Research article

In vitro studies of amyloid β-protein fibril assembly and toxicity provide clues to the aetiology of Flemish variant (Ala692→Gly) Alzheimer's disease

Dominic M. WALSH, Dean M. HARTLEY, Margaret M. CONDRON, Dennis J. SELKOE, David B. TEPLOW

Abstract

In a Flemish kindred, an Ala692 → Gly amino acid substitution in the amyloid β-protein precursor (AβPP) causes a form of early-onset Alzheimer's disease (AD) which displays prominent amyloid angiopathy and unusually large senile plaque cores. The mechanistic basis of this Flemish form of AD is unknown. Previous in vitro studies of amyloid β-protein (Aβ) production in HEK-293 cells transfected with cDNA encoding Flemish AβPP have shown that full-length [Aβ(1–40)] and truncated [Aβ(5–40) and Aβ(11–40)] forms of Aβ are produced. In an effort to determine how these peptides might contribute to the pathogenesis of the Flemish disease, comparative biophysical and neurotoxicity studies were performed on wild-type and Flemish Aβ(1–40), Aβ(5–40) and Aβ(11–40). The results revealed that the Flemish amino acid substitution increased the solubility of each form of peptide, decreased the rate of formation of thioflavin-T-positive assemblies, and increased the SDS-stability of peptide oligomers. Although the kinetics of peptide assembly were altered by the Ala21 → Gly substitution, all three Flemish variants formed fibrils, as did the wild-type peptides. Importantly, toxicity studies using cultured primary rat cortical cells showed that the Flemish assemblies were as potent a neurotoxin as were the wild-type assemblies. Our results are consistent with a pathogenetic process in which conformational changes in Aβ induced by the Ala21 → Gly substitution would facilitate peptide adherence to the vascular endothelium, creating nidi for amyloid growth. Increased peptide solubility and assembly stability would favour formation of larger deposits and inhibit their elimination. In addition, increased concentrations of neurotoxic assemblies would accelerate neuronal injury and death.

  • fibrillogenesis
  • neurotoxicity